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Previous studies overlappingly used Barbonymus balleroides, Systomus orphoides, and Systomus rubripinnis as scientific names for three barb species living in Klawing River, i.e., for 'brek', 'lukas', and 'maracoca' fish. This taxonomic chaos can be solved by applying DNA barcoding, using the cytochrome c oxidase 1 gene for precise species identification. This study evaluated the taxonomic status of barb species (Cyprinidae) living in Klawing River, Purbalingga Regency, Central Java, Indonesia. Fish samples were randomly collected from the upper stream to downstream using spread nets targeting consumable adult individuals. Pectoral fin clips were cut off using scissors and preserved in ethanol 96%. Morphological identification was carried out by comparing the fish samples to images available in references. The genetic identity was determined based on similarity, homology tests, and monophyly to reference species in databases. The current study genetically identified barb species from Klawing River as Labiobarbus fasciatus (100% and 100%) for lukas, Barbonymus schwanefeldii (>96.80 and >96.80) for brek, and Leptobarbus melanopterus (>99.80 and >99.80) for maracoca. There was high genetic identity, genetic similarity, and monophyly with their conspecific references in BOLD systems and GenBank, respectively. None of the barb samples were identified as B. balleroides, S. orphoides, or S. rubripinnis, as previously utilized scientifically for the three barb species from the Klawing River. Instead of validating B. balleroides, S. orphoides, and S. rubripinnis, as previously employed scientific names for barb species, this research alleged possible misidentification of barb species and validates that the barb fishes in Klawing River, Central Java, Indonesia belong to B. schwanefeldii, L. fasciatus, and Leptobarbus melanopterus.